This invention relates to type IV collagenase, hereinafter also referred to as gelatinase. More particularly, the invention relates to the cDNA clone representing the full size human type IV collagenase (gelatinase).
Collagens constitute the most abundant proteins of the extracellular matrix (ECM) in mammalian organisms. Collagen and other macromolecules of the ECM are deposited by resident cells and organized into a three-dimensional meshwork. This ECM environment plays an essential role in guiding cell migration, and in cell-to-cell communication during morphogenetic processes. The restructuring of the ECM during remodeling occurs as a cooperative multistep process involving a localized degradation of existing macromolecules, rearrangement of the cytoskeleton, cell translocation, and deposition of new ECM components.
The few secreted proteases capable of initiaing the degradation of ECM proteins previously identified include; fibroblast and granulocyte collagenases which degrade interstitial collagens, collagenase degrading type IV basement membrane collagen, stromelysin and gelatinase. Examination of the specific role of each protease in ECM metabolism is complicated by the difficulty of differential identification of the enzymes.
Type IV collagenase (gelatinase) represents a new member of an emerging gene family coding for secreted ECM metalloproteases. Initial identification, purification and/or partial characterization of this protease is described by Seltzer et al., J. Biol. Chem. 256, 4662-4668 (1981); Sopata, Biochim. Biophys. Acta 717, 26-31 (1982); Seltzer et al., J. Chromatog. 326, 147-155 (1985); Murphy et al., Biochim. Biophys. Acta 831, 49-58 (1985); and Hibbs et al., J. Biol. Chem. 260, 2493-2500 (1985).
Recent advances in biochemistry and in recombinant DNA technology have made it possible to synthesize specific proteins, for example, enzymes, under controlled conditions independent of the organism from which they are normally isolated. These biochemical synthetic methods employ enzymes and subcellular components of the protein synthesizing systems of living cells, either in vitro in cell-free systems, or in vivo in microorganisms. In either case, the principal element is provision of a deoxyribonucleic acid (DNA) of specific sequence which contains the information required to specify the desired amino acid sequence. Such a specific DNA sequence is termed a gene. The coding relationships whereby a deoxyribunucleotide sequence is used to specify the amino acid sequence of a protein is well-known and operates according to a fundamental set of principles. See, for example, Watson, Molecular Biology of the Gene, 3d ed., Benjamin-Cummings, Menlo Park, Calif., 1976.
A cloned gene may be used to specify the amino acid sequence of proteins synthesized by in vitro systems. RNA-directed protein synthesizing systems are well-established in the art. Doublestranded DNA can be induced to generate messenger RNA (mRNA) in vitro with subsequent high fidelity translation of the RNA sequence into protein.
It is now possible to isolate specific genes or portions thereof from higher organisms, such as man and animals, and to transfer the genes or fragments to microorganisms such as bacteria or yeasts. The transferred gene is replicated and propogated as the transformed microorganism replicates. Consequently, the transformed microorganism is endowed with the capacity to make the desired protein or gene which it encodes, for example, an enzyme, and then passes on this capability to its progeny. See, for example, Cohen and Boyer, U.S. Pat. Nos. 4,237,224 and 4,468,464.